A variety of diseases including asthma, cancers, heart diseases, aneurysms, autoimmune diseases, and viral infections manifest varying symptoms and signs and yet it has been suggested that an abnormal expression (an overexpression or underexpression) of one or a few proteins is a major etiologic factor in many cases. In general, the expression of those proteins is controlled by a variety of transcriptional regulatory factors such as transcription activating factors and transcription suppressing genes.
NF-κB is a transcriptional regulatory factor consisting of heterodimers p65 and p50. NF-κB is typically localized in the cytoplasm where NF-κB is bound by its inhibitory factor Iκ so that intranuclear movement of NF-κB is prevented. However, when a stimulus, such as cytokine, ischemia, reperfusion, or the like, is applied due to any cause, IκB is degraded by phosphorylation. As a result, NF-κB is activated and transferred into the nucleus. In the nucleus, NF-κB binds to an NF-κB binding site on a chromosome and promotes the transcription of a gene downstream thereof. As genes located downstream of the NF-κB binding site, for example, inflammatory cytokines (e.g., IL-1, IL-6, IL-8, tumor necrosis factor α (INF α), etc.) and adhesion molecules (e.g., (e.g., VCAM-1, ICAM-1, etc.) are known.
NF-κB may be involved in the onset of progression of tumor malignancy (Rayet B et al., Oncogene 1999 Nov. 22; 18(49)6938-47); NF-κB is involved in response of tumor cells to hypoxia stress (Royds J A et al., Mol Pathol 1998 April; 51(2):55-61); NF-κB inhibits expression of cytokines and adhesion molecules in synovial membrane cells derived from chronic rheumatoid arthritis patients (Tomita T et al., Rheumatology (Oxford) 2000 July; 39(7):749-57); suppression of coordination between a plurality of transcriptional factors including NF-κB changes the malignant phenotypes of various tumors (Denhardt D. T., Crit. Rev. Oncog., 1996; 7(3-4):261-91); downregulation of NF-κB activation due to green tea polyphenol blocks induction of nitric oxide synthesizing enzyme, and suppresses A431 human epidermoid carcinoma cells (Lin J. K., et al., Biochem. Pharmacol., 1999, Sep. 15; 58(6):911-5); amyloid β peptide observed in the brains of Alzheimer's disease patients binds to 75-kD neurotrophic receptor (p75NTR) in neuroblastoma cells to activate NF-κB in a time-dependent manner and a dose-dependent manner (Kuper P, et al., J. Neurosci. Res., 1998, Dec. 15; 54(6):798-804); TNF-α, which is activated by NF-κB, plays an important role in the onset of glomerulonephritis (Ardaillou et al., Bull. Acad. Natl. Med., 1995, Jan; 179(1)103-15); NF-κB decoy in vivo blocks expression of cytokines and adhesion molecules in mouse nephritis induced by TNF α (Tomita N., et al., Gene Ther., 2000, Aug.; 7(15)1326-32); and the like.
It was suggested that NF-κB suppresses MMP1 and MMP9, members of matrix metalloproteinase (MMP), at the transcription level (Eberhardt W., Huwiler A., Beck K. F., Walpen S., Pfeilschifter J., “Amplification of IL-1β-induced matrix metalloproteinase-9 expression by superoxide in rat glomerular mesangial cells is mediated by increased activities of NF-κB and activating protein-1 and involves activation of the mitogen-activated protein kinase pathways”, J. Immunol., 2000, Nov. 15, 165(10), 5788-97; Bond M., Baker A. H., Newby A. C., “Nuclear factor KB activity is essential for matrix metalloproteinase-1 and -3 upregulation in rabbit dermal fibroblasts”, Biochem. Biophys. Res. Commun., 1999, Oct. 22, 264 (2), 561-7; Bond M., Fabunmi R. P., Baker A. H., Newby A. C., “Synergistic upregulation of metalloproteinase-9 by growth factors and inflammatory cytokines: an absolute requirement for transcriptional factor NF-κB”, FEBS Lett., 1998, Sep. 11, 435(1), 29-34; and Kim H., Koh G., “Lipopolysaccharide activates matrix metalloproteinase-2 in endothelial cells through an NF-κB-dependent pathway”, Biochem. Biophys. Res. Commun., 2000, Mar. 16, 269(2), 401-5).
MMP is a polygene family of zinc-dependent enzymes involved in degradation of extracellular matrix components. It is also known that ets suppresses MMP1 and MMP9, members of matrix metalloproteinase (MMP), at the transcription level (Sato Y., Abe M., Tanaka K., Iwasaka C., Oda N., Kanno S., Oikawa M., Nakano T., Igarashi T., “Signal transduction and transcriptional regulation of angiogenesis”, Adv. Exp. Med. Biol., 2000, 476, 109-15; and Oda N., Abe M., Sato Y., “ETS-1 converts endothelial cells to the angiogenic phenotype by inducing the expression of matrix metalloproteinases and integrin β3”, J. Cell Physiol., 1999, February, 178 (2), 121-32).
MMP plays an important role in invasion of cancer cells by mediating degradation of extracellular matrix protein. A number of studies suggested the involvement of MMP and MMP inhibitors (TIMP) in the progression of cancer: the TIMP1 level in serum may be used as a marker for prognosis and diagnosis of colon and rectum cancer, and as a selective marker for metastatic cancer (Pellegrinl P., et al., Cancer Immunol. Immunother., 2000 September; 49 (7):388-94); expression and activity of MMP2 and MMP9 in human urinary bladder cancer cells are affected by tumor necrosis factor α and γ interferon (Shin K Y et al., Cancer Lett 2000 Oct. 31; 159 (2):127-134); MMP2, MMP9 and MT1-MMP, and their inhibitors, TIMP1 and TIMP2, are expressed in ovarian epithelium tumor (Sakata K., et al., Int. J. Oncol., 2000, Oct.; 17 (4):673-681); the level of each of MMP1, MMP2, MMP3 and MMP9 and the overall MMP activity are upregulated in colon and rectum tumor, and MMP1 is most important for progression of colon and rectum cancer (Baker E. A., et al., Br. J. Surg., 2000, September; 87(9):1215-1221); activated MMP2 plays an important role in invasion of urothelial cancer, and also the expression level of the activated MMP2 can be used as a useful prognosis index (KanedaK., et al., BJU Int., 2000, September; 86(4):553-557); a prostaglandin synthesis inhibitor inhibits invasion of human prostate tumor cells, and reduces the release of MMP (Attiga F. A., et al., Cancer Res., 2000, Aug. 15; 60(16):4629-37); the MMP activity of a serum euglobulin fraction increases in breast cancer and lung cancer patients, and may be used as a tumor marker for these cancers (Farias E., et al., Int. J. Cancer, 2000, Jul. 20; 89(4):389-94); a MMP inhibitor inhibits gelatin-degrading activity in tumor cells (Ikeda M., et al., Clin. Cancer Res., 2000, Aug.; 6(8):3290-6); induction of MMP9 due to a membrane protein LMP1 contributes to metastatic of nasopharyngeal cancer (NPC) (Horikawa T., et al., Cancer, 2000, Aug. 15; 89(4):715-23); MMP plays an important role in an early stage of angioplasty, and a MMP inhibitor suppresses invasion and morphogenesis of human microvascular endothelial cells (Jia M. C., et al., Adv. Exp. Med. Biol., 2000; 476:181-94); MMP9 is expressed in invasive and recurrent pituitary adenoma and hypophysis cancer (Turner H. E., et al., J. Clin. Endocrinol. Metab., 2000, August; 85(8):2931-5); and the like.
MMP is also known to be involved in development of aortic aneurysm: MMP is involved in formation and rupture of cerebral aneurysm (Gaetani P., et al., Neurol. Res., 1999, Jun.; 21(4):385-90); a MMP-9 promotor is a risk factor for cerebral aneurysm (Peters D. G., et al., Stroke, 1999, December; 30(12):2612-6); inhibition of MMP inhibits the growth of microaneurysm in an aneurysm model (Treharne G. D., et al., Br. J. Surg., 1999, August; 86(8):1053-8); and the like.
MMP is secreted from migrating vascular smooth muscle cells, macrophage, and the like, and destroys collagen, elastin, and the like present in blood vessel walls, whereby the tension of the blood vessel is lost and the blood vessel does not resist the blood pressure and its diameter is expanded. In fact, in the blood vessel of an aneurysm, significant destruction of elastin is observed (Halloran B. G., Baxter B. T., “Pathogenesis of aneurysms”, Semin. Vasc. Surg., 1995, Jun. 8, (2):85-92).
Aortic aneurysmal rupture is substantially fatal. To prevent aortic aneurysmal rupture, it is important to remove risk factors of arteriosclerosis. However, it is difficult to eliminate the risk factors. At present, invasive surgery is the only means for preventing aortic aneurysmal rupture.
According to data obtained by measuring the aorta diameter of from 35-year-old to 80-year old adult males, the average was 1.5 cm to 2.0 cm (Dolores J Katz, James C. Stanley, Gerald B. Zelenock, “Abdominal Aortic Aneurysms”, Seminars in Vascular Surgery, vol. 8, No. 4 (December), 1995; pp. 289-298). In general, the aorta having a diameter beyond 1.5 times as great as the average value is judged as an aortic aneurysm. However, according to the above-described data, one in every 400 people had an aneurysm having a diameter of 3 cm or more which is judged as aortic aneurysm. Therefore, although the degree of risk of aorta rupture is not considered here, the prevalence of aortic aneurysm is relatively high in from 35-year-old to 80-year old adult males. The prevalence is believed to be even greater in males aged 65 and above.
It is known that MMPs are involved in chronic articular rheumatism: alleviation of chronic articular rheumatism by drug treatment leads to a decrease in MMP1 within synovial membrane tissue (Kraan M. C., et al., Arthritis Rheum., 2000, August; 43(8):1820-30); upregulation of MT-MMP expression by IL-1β partially induces activation of MMP-2, leading to cytokine-mediated articular disruption in chronic articular rheumatism (Origuchi T., Clin. Exp. Rheumatol., 2000, May-Jun.; 18(3):333-9); inflammatory cytokine IL-17 produced in synovial membrane of chronic articular rheumatism increases production of MMP1 (Chabaud M., et al., Cytokine, 2000, Jul.; 12(7):1092-9); MMP1, MMP2, MMP3, MMP8, MMP9 and an MMP inhibitor are present in the chronic articular rheumatism synovia at a high level, when MMPs are activated, the balance with the MMP inhibitor is lost, resulting in cartilage disruption (Yoshihara Y., et al., Ann. Rheum. Dis., 2000, June; 59(6):455-61); MT1-MMP is involved in activation of proMMP-2 in rheumatic synovial membrane lining cell layers, leading to cartilage disruption in chronic articular rheumatism (Yamanaka H., et al., Lab. Invest., 2000, May; 80(5):677-87); and the like.
MMP is involved in cardiovascular lesions due to Marfan's syndrome (Segura A. M., et al., Circulation, 1998, Nov. 10; 98(19 Suppl):11331-7).
Expression of membrane type MMP (MT-MMP) is increased in mesangial proliferative glomerulonephritis (Hayashi K., et al., J. Am. Soc. Nephroi., 1998, Dec.; 9(12):2262-71).
It has been reported that a MMP inhibitor suppresses the expansion of a blood vessel diameter in an aortic aneurysm model in rat abdomen (Moore G., Liao S., Curci J. A., Starcher B. C., Martin R. L., Hendricks R. T., Chen J. J., Thompson R. W., “Suppression of experimental abdominal aortic aneurysms by systemic treatment with a hydroxamate-based matrix metalloproteinase inhibitor” (RS 132908), J. Vasc. Surg., 1999, Mar.; 29(3):522-32).
A MMP inhibitor may be used in therapy for glomerulonephritis (Marti H P, Schweiz Med Wochenschr 2000 May 27; 130(21); 784-8). However, systemic administration of a MMP inhibitor causes severe side effects, and has difficulty in clinical applications for treatment (therapy and prevention) of various diseases.
It has been suggested that NF-κB is involved in various diseases via expression of a number of genes under the transcription control thereof. However, no method for effectively treating these diseases, particularly a non-invasive treatment method, has been provided. Particularly, as described above, aortic aneurysm is not a rare disease. As society ages, an increase in arteriosclerotic diseases inevitably leads to an increase in aortic aneurysm diseases. Considering the aging of patients, it is ideal to suppress directly the growth of aortic aneurysm using a pharmaceutical agent, however, to date such a means is not present. There is a desperate demand for development of a low-invasive therapy and prevention method for aortic aneurysm.